Thinking of Co²⁺-staining explant tissue or cultured cells? How to make it reliable and specific

Ca(2+) and/or Zn(2+) entry into neurons and glial cells is often a key step driving the processes of neurodevelopment and disease. As a result, a major pre-occupation of many neuroscientists has been in tracking down when and where nervous tissues express ion channels with appreciable divalent ion permeability. The cobalt (Co(2+))-staining technique is one of the few techniques that allow a snapshot of the entire neuronal circuit, and selectively labels cells expressing divalent-permeable ion channels with a brown-black precipitate. Despite this, its use has been remarkably limited in the past decade. Reluctance to employ this approach has largely been related to an earlier concern with obtaining a reliable and reproducible means of visualizing transported Co(2+). Here we show that recent advances have resolved these issues, opening this straightforward and valuable technique to a much larger neuroscience audience.